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. 2017 Aug 8;7:7594. doi: 10.1038/s41598-017-07380-9

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© The Author(s) 2017

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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An inducible system for AID expression and accumulation in the nucleus. (A) Representative scheme of the HA-tagged AID retroviral construct. (B) AID expression is induced following doxycycline (D) treatment and retained in the nucleus upon additional treatment with leptomycin B (L), which specifically inhibits nuclear export. Anti-HA antibody was used to detect AID expression and histone H3 was used as a loading control (C) Representative confocal images of AID subcellular localisation in HeLa and Jiyoye cells after doxycycline and leptomycin B treatment. Nuclear DNA was counterstained with DAPI (red). A total of 25 cells from randomly selected fields were analysed in each experimental condition. The graphs in the right show the quantification of the cellular signal of AID within the cells. Light gray section of the bar indicates the average percentage of cytoplasmic AID signal. Black section of the bar indicates the average percentage of nuclear AID signal. (D) Association of AID with heterochromatin. Time course digestion of HA-tagged AID Jiyoye cells nuclei with DNase I, in which the supernatant contains the euchromatin fraction (as demonstrated by the appearance of H3Ac and H3K4me3) and the pellet contains the heterochromatin fraction (as shown by the progressive decrease of H4K20me3 and H3K27me3).